Single-Molecule Studies of Allosteric Inhibition of Individual Enzyme on a DNA Origami Reactor
文献类型：期刊论文
| 作者 | Xu, Y; Gao, YJ; Su, YY; Sun, LL; Xing, FF; Fan, CH; Li, D |
| 刊名 | JOURNAL OF PHYSICAL CHEMISTRY LETTERS
 |
| 出版日期 | 2018 |
| 卷号 | 9期号:23页码:6786-6794 |
| 关键词 | HORSERADISH-PEROXIDASE SUPERRESOLUTION MICROSCOPY CONFORMATIONAL DYNAMICS DISTAL HISTIDINE RESONANCE RAMAN ACTIVE-SITE BINDING MECHANISM CATALYSIS KINETICS |
| ISSN号 | 1948-7185 |
| DOI | 10.1021/acs.jpclett.8b02992 |
| 文献子类 | 期刊论文 |
| 英文摘要 | Unraveling the conformational changes of enzymes together with inhibition kinetics during an enzymatic reaction has great potential in screening therapeutic candidates; however, it remains challenging due to the transient nature of each intermediate step. We report our study on the noncompetitive inhibition of horseradish peroxidase with single-turnover resolution using single-molecule fluorescence microscopy. By introducing DNA origami as an addressable nanoreactor, we observe the coexistence of nascent-formed fluorescent product on both catalytic and docking sites. We further propose a single-molecule kinetic model to reveal the interplay between product generation and noncompetitive inhibition and find three distinct inhibitor releasing pathways. Moreover, the kinetic isotope effect experiment indicates a strong correlation between catalytic and docking sites, suggesting an allosteric conformational change in noncompetitive inhibition. A memory effect is also observed. This work provides an in-depth understanding of the correlation between enzyme behavior and enzymatic conformational fluctuation, substrate conversion, and product releasing pathway and kinetics. |
| 语种 | 英语 |
| 源URL | [http://ir.sinap.ac.cn/handle/331007/31110]  |
| 专题 | 上海应用物理研究所_中科院上海应用物理研究所2011-2017年 |
| 作者单位 | 1.Natl Engn Res Ctr Nanotechnol, Shanghai 200241, Peoples R China 2.Chinese Acad Sci, Shanghai Inst Appl Phys, Key Lab Interfacial Phys & Technol, Bioimaging Ctr,Shanghai Synchrotron Radiat Facil, Shanghai 201800, Peoples R China 3.Univ Chinese Acad Sci, Beijing 100049, Peoples R China 4.Shanghai Univ, Coll Sci, Dept Chem, Shanghai 200444, Peoples R China 5.East China Normal Univ, Sch Chem & Mol Engn, Shanghai 200241, Peoples R China 6.Chinese Acad Sci, Shanghai Inst Appl Phys, Div Phys Biol, Shanghai 201800, Peoples R China |
| 推荐引用方式 GB/T 7714 | Xu, Y,Gao, YJ,Su, YY,et al. Single-Molecule Studies of Allosteric Inhibition of Individual Enzyme on a DNA Origami Reactor[J]. JOURNAL OF PHYSICAL CHEMISTRY LETTERS,2018,9(23):6786-6794. |
| APA | Xu, Y.,Gao, YJ.,Su, YY.,Sun, LL.,Xing, FF.,...&Li, D.(2018).Single-Molecule Studies of Allosteric Inhibition of Individual Enzyme on a DNA Origami Reactor.JOURNAL OF PHYSICAL CHEMISTRY LETTERS,9(23),6786-6794. |
| MLA | Xu, Y,et al."Single-Molecule Studies of Allosteric Inhibition of Individual Enzyme on a DNA Origami Reactor".JOURNAL OF PHYSICAL CHEMISTRY LETTERS 9.23(2018):6786-6794. |
入库方式： OAI收割
来源：上海应用物理研究所
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